用于原位联动检测二硫化碳诱发活性小分子协同调控细胞信号转导的荧光探针

In comparison with other biological detection technologies, fluorescence bioimaging technology has become a powerful supporting tool, which provides an attractive technique to study physiological and pathological processes of interest with high spatial and temporal resolution, less invasiveness, and more convenience. Contrapose the existing problem that whether carbon disulfide is able to induce changes of endogenous bioactive species and regulate the cellular signal transduction or not, this project is proposed to design and screen new fluorescence probes and fluorescence labels with new techniques for three - dimensional tracing and high spatial and temporal resolution imaging on the levels of cells, tissues and livings so that the interactive detection of endogenous bioactive species which induced by CS2 will be presented visually, in-real time and in situ. In order to achieve the real-time, dynamic, high-throughput interactive detection for variety targets in vivo, the fluorescence interaction imaging system with high-throughput detection capability should be constructed by integrating the advantages of fluorescence probes and fluorescence labels for bioimaging. It is expected to illustrate the distribution and transportation of CS2 in vivo and investigate the rules of the interaction between CS2 and numerous endogenous species with the imaging analysis method based on the fluorescence probes and fluorescence labels. This project will not only contribute to promote the development of fluorescence imaging analysis, also provide powerful technique supports for the solution to the problem of the environmental toxicity of CS2 and our health, and the exploration of cell signal transduction and physiological function pathway induced by CS2 from the inside and the outside.

荧光探针成像技术以其时空分辨率高、操作简便和原位非损伤检测等优点，在生物活性物种检测领域，已成为一种功能强大的研究辅助工具。本项目针对二硫化碳(CS2)是否可以诱导生物内源活性物种变化来调控细胞信号通路的高灵敏联动检测的难题，拟设计和筛选出新型荧光探针和荧光标记，利用探针的三维示踪和高时空分辨成像新技术，在细胞、组织和活体水平上，实现CS2诱导下生物内源活性分子的原位、实时、可视化联动检测。整合荧光探针和荧光标记所具有的生物成像技术优势，构建具有高通量检测能力的荧光联动成像系统，实现多种靶标物质在活体内的实时、动态、高通量联动检测。期冀借助基于荧光探针和荧光标记建立的成像分析方法，阐明CS2在生命体内的分布、转运以及与内源物种相互作用的规律。本项目将有助于推动荧光成像分析的发展，为解决CS2调控生理作用的深层次问题，为明晰内外源性CS2诱导下细胞信号转导和生理功能通路等提供有力的技术支撑。

荧光生物成像技术是一种新型生物分析检测平台。通过设计光学探针标记特定研究对象，利用多光学成像模式原位、实时、动态地在分子、细胞、组织和活体水平上定性和定量地获取分析对象的体内相关信息。鉴于CS2潜在调控细胞信号转导通路的重要生物医学意义，发展用于协同检测CS2协同相关活性物种变化水平的分析方法已经迫在眉睫。本项目针对CS2诱导生物内源活性物种变化的高灵敏可视化分析的难题，拟设计和筛选出新型荧光探针，利用荧光探针的三维示踪和高时空分辨成像新技术，在细胞、组织和活体水平上，实现对内源/外源CS2及其诱导下生物内源活性分子的原位、实时、可视化联动检测。本项目执行期间在高水平国际学术期刊Angewandte Chemie、Biomaterials、ACS Sensors、Sensors & Actuators: B. Chemical、Analytical Chemistry发表论文14篇，授权中国发明专利2项。.1）近红外比率型荧光探针成像分析CS2胁迫下细胞内硒醇浓度变化。.2）检测CS2刺激谷胱甘肽变化的双光子荧光探针在喉癌手术导航中的潜在应用。.3）CS2调控肿瘤细胞死亡的信号通路研究。.4）CS2潜在酶调控肿瘤细胞死亡的信号通路的前期研究。.5）细胞内与CS2有潜在联系的活性物种的检测与前期研究。